Konfokales Live-Imaging Mikroskop mit Spinning Disc
Final Report Abstract
The spinnig disc was intensively used for live imaging experiments as well as for microscopy on fixed tissue samples. In live imaging experiments we used both dissociate neurons and brain slices. In one set of experiments we studied molecular mechanisms of growth and dynamics of inner cytoskeletal scaffold formed by microtubules during axon navigation in vivo. We discovered that that Sip1 transcription factor control the growth rate of axonal MTs through ninein. To assess this, we used the MT plus-end binding protein EB3 fused with GFP and tracked GFP-EB3 comets through live-imaging experiments. The average anterograde velocity of GFP-EB3 comets was significantly reduced in Sip1- deficient neurons. When we examined axons of Sip1-deficient neurons where ninein expression was restored, we observed that the average anterograde velocity of GFP- EB3 comets was significantly higher than in the axons of Sip1-deficient neurons. In order to measure the microtubule dependent velocity of GFP-EB3 comets, we tracked them for 10 s or more. We used the microscope to record the mitochondrial dynamic via life-cell-imaging in human primary fibroblasts obtained from patients with mitochondriopathy. The microscope was used in a study investigating the influence of systemic Propionylmannosamine application using a specific in vivo mouse model. Using mice expressing axonal fluorescent proteins, we quantified the extension of regenerating axons, the number of regenerating axons, the number of arborising axons and the number of branches per axon 5 days after injury. Spine morphology in NOMA-GAP mutants was investigated. Axon growth in Satb2 and Ctip2 mutants was investigated. The role of NT3 in the cell fate of neocortical progenitors was studied.
Publications
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Ntf3 acts downstream of Sip1 in cortical postmitotic neurons to control progenitor cell fate through feedback signaling. Development. 2014 Sep;141(17):3324-30
Parthasarathy S, Srivatsa S, Nityanandam A, Tarabykin V
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Bcl11a (Ctip1) Controls Migration of Cortical Projection Neurons through Regulation of Sema3c. Neuron. 2015 Jul 15;87(2):311-25
Wiegreffe C, Simon R, Peschkes K, Kling C, Strehle M, Cheng J, Srivatsa S, Liu P, Jenkins NA, Copeland NG, Tarabykin V, Britsch S
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miR-128 regulates neuronal migration, outgrowth and intrinsic excitability via the intellectual disability gene Phf6. Elife. 2015 Jan 3;4
Franzoni E, Booker SA, Parthasarathy S, Rehfeld F, Grosser S, Srivatsa S, Fuchs HR, Tarabykin V, Vida I, Wulczyn FG
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N-Propionylmannosamine stimulates axonal elongation in a murine model of sciatic nerve injury. Neural Regen Res. 2015 Jun;10(6):976-81
Witzel C, Reutter W, Stark GB, Koulaxouzidis G
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NOMA-GAP/ARHGAP33 regulates synapse development and autistic-like behavior in the mouse. Mol Psychiatry. 2015 Sep;20(9):1120-31
Schuster S, Rivalan M, Strauss U, Stoenica L, Trimbuch T, Rademacher N, Parthasarathy S, Lajkó D, Rosenmund C, Shoichet SA, Winter Y, Tarabykin V, Rosário M
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Sip1 downstream Effector ninein controls neocortical axonal growth, ipsilateral branching, and microtubule growth and stability. Neuron. 2015 Mar 4;85(5):998-1012
Srivatsa S, Parthasarathy S, Molnár Z, Tarabykin V
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Homozygous YME1L1 mutation causes mitochondriopathy with optic atrophy and mitochondrial network fragmentation; Elife. 2016 Aug 6;5. pii: e16078
Hartmann et al.
